title: Silencing subtelomeric VSGs by Trypanosoma brucei RAP1 at the insect stage involves chromatin structure changes.
abstract: Trypanosoma brucei causes human African trypanosomiasis and regularly switches its major surface antigen variant surface glycoprotein (VSG) to evade mammalian host immune responses at the bloodstream form (BF) stage. Monoallelic expression of BF Expression Site (BES)-linked VSGs and silencing of metacyclic VSGs (mVSGs) in BF cells are essential for antigenic variation, whereas silencing of both BES-linked and mVSGs in the procyclic form (PF) cells is important for cell survival in the midgut of its insect vector. We have previously shown that silencing BES-linked VSGs in BF cells depends on TbRAP1. We now show that TbRAP1 silences both BES-linked and mVSGs at both BF and PF stages. The strength of TbRAP1-mediated BES-linked VSG silencing is stronger in the PF cells than that in BF cells. In addition, Formaldehyde-Assisted Isolation of Regulatory Elements analysis and MNase digestion demonstrated that depletion of TbRAP1 in PF cells led to a chromatin structure change, which is significantly stronger at the subtelomeric VSG loci than at chromosome internal loci. On the contrary, no significant chromatin structure changes were detected on depletion of TbRAP1 in BF cells. Our observations indicate that TbRAP1 helps to determine the chromatin structure at the insect stage, which likely contributes to its strong silencing effect on VSGs.
authors.affiliation: Department of Biological, Geological, and Environmental Sciences, Center for Gene Regulation in Health and Disease, Cleveland State University, Cleveland, OH 44115, USA.
authors.forename: Unnati M
authors.initials: UM
authors.lastname: Pandya
authors.forename: Ranjodh
authors.initials: R
authors.lastname: Sandhu
authors.forename: Bibo
authors.initials: B
authors.lastname: Li
mesh_terms: D002843:Chromatin / Q000737:chemistry*; D020868:Gene Silencing; D015800:Protozoan Proteins / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D016615:Telomere; D014346:Trypanosoma brucei brucei / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000235:genetics* / Q000378:metabolism
chemical_list: D002843:Chromatin; D015800:Protozoan Proteins; D012333:RNA, Messenger; D014643:Variant Surface Glycoproteins, Trypanosoma